Mechanism of miR-107/HMOX1 axis in hepatic sinusoidal endothelial cells stimulated by ischemia-reperfusion injury

Abstract

Background: Liver ischemia-reperfusion injury (IRI) is a common complication of diseases such as liver transplantation, hepatic resection, and hemorrhagic shock. This study aimed to elucidate the molecular mechanism of miR-107 affecting hepatic ischemia-reperfusion injury (IRI).

Methods: The expression changes of miR-107 during hepatic IRI were quantified using quantitative real-time PCR. Subsequently, in vitro cellular experiments were carried out to verify the role of miR-107 on hypoxia/reoxygenation (HR) through CCK-8, flow cytometer, and commercial kits. In terms of mechanism, it was determined that miR-107 had a regulatory relationship with target genes through luciferase reporter assay.

Results: In mouse liver IRI, miR-107 expression was increased while HMOX1 expression was decreased in liver tissues. In vitro cellular experiments, miR-107 inhibitors favored the alleviation of proliferation, apoptosis, inflammation, and oxidative stress in HR-damaged liver sinusoidal endothelial cells. In the molecular mechanism study, we determined that miR-107 could bind to HMOX1 and inhibit the HMOX1 expression. Low HMOX1 expression could eliminate the protective effect of miR-107 inhibitors.

Conclusion: MiR-107 expression was elevated during hepatic IRI and exacerbates hepatic injury by targeting HMOX1 inhibition.

Keywords: HMOX1; Hepatic IRI; Inflammation; Oxidation; miR-107.

Fig. 1

The experimental results graph of hepatic IRI vividly illustrates the pathological progression of liver damage by comparing key metrics at distinct time points (sham operation, IRI 3 days, 5 days, 7 days). (A) The general appearance of the liver (visual observation): As time progresses, the liver exhibits increasing signs of congestion, swelling, and localized necrosis (e.g., dark purple regions), which suggests that ischemia-reperfusion injury (IRI) induces pronounced morphological alterations in the liver. The severity of damage appears to peak between 3 and 5 days. (B) HE staining (histopathological morphology, light microscopy observation) confirmed that ischemia-reperfusion injury (IRI) induced significant pathological damage to liver cells. (C) The results of tissue immunofluorescence LYVE-1 labeling of LSECs. (D) Tissue immunofluorescence labeling with LYVE-1 and PCNA. (E) Immunohistochemistry results using PCNA

Fig. 2

The IRI treatment induced the upregulation of miR-107 expression. **P < 0.01, ***P < 0.001, compared to sham group

Fig. 3

Significance of miR-107 on LSECs. (A) H/R treatment induced the significant upregulation of miR-107, and its inhibitor could effectively suppress this upregulation. (B) Proliferative ability was recovered after silencing miR-107. (C) Knockdown of miR-107 reversed the abnormal apoptosis in the HR group. (D-E) Protective roles of silenced miR-107 on inflammation and oxidation. ***P < 0.001, compared to control group; ###P < 0.001, compared to HR group

Fig. 4

miR-107 regulated HMOX1 expression. (A) The target binding sites of miR-107 and HMOX1. (B) The dual-luciferase reporter assay was employed to validate the direct targeting relationship between miR-107 and HMOX1. (C) miR-107 negatively regulated the expression level of HMOX1. (D) The expression levels of HMOX1 induced by IRI. ***P < 0.001, compared to the control group or sham group

Fig. 5

The miR-107/HMOX1 axis plays a regulatory role in LSECs. (A) Regulation of HMOX1 levels under different treatments. (B-E) miR-107 modulates proliferation, apoptosis, inflammation, and oxidation in LSECs by directly targeting HMOX1. ***P < 0.001, compared to control group; ###P < 0.001, compared to HR group; &P < 0.05, &&&P < 0.001, compared to HR + miR-inhibitor group